胡万福 刘明 杨燕 李玲 李广松

摘 要 目的：探討清肺保元胶囊对慢性阻塞性肺疾病（COPD）模型大鼠气道炎症的抑制作用，以及对NOD样受体蛋白3（NLRP3）信号通路的影响。方法：将60只SD雄性大鼠随机分为空白对照组、模型组、地塞米松组（阳性对照，0.2 mg/kg）和清肺保元胶囊高、中、低剂量组（1 232.0、616.0、308.0 mg/kg），每组10只。除空白对照组外，其余各组大鼠经烟熏28 d和气管内滴注脂多糖2次以复制COPD模型。自实验第29天起，空白对照组和模型组大鼠灌胃等体积生理盐水，各给药组大鼠灌胃相应药物;给药体积均为10 mL/kg，每日1次，连续28 d。末次给药后，检测各组大鼠肺功能;采用苏木精-伊红染色法观察其肺组织病理学变化，采用酶联免疫吸附测定法检测其支气管肺泡灌洗液中白细胞介素1β（IL-1β）的含量并进行白细胞计数;采用Western blotting法检测其肺组织中NLRP3、剪切的胱天蛋白酶1（Cleaved caspase-1）蛋白的相对表达量。结果：模型组、地塞米松组和清肺保元胶囊高、中、低剂量组分别有3、2、1、1、2只大鼠死亡。与空白对照组比较，模型组大鼠第0.3秒用力呼气容积/用力肺活量比值（FEV0.3/FVC）显著降低（P<0.01）;其肺组织可见大量炎性细胞浸润，病变明显;其支气管肺泡灌洗液中IL-1β含量和白细胞计数以及肺组织中NLRP3、Cleaved caspase-1蛋白的相对表达量均显著升高（P<0.05或P<0.01）。与模型组比较，各给药组大鼠FEV0.3/FVC均显著升高（P<0.05或P<0.01）;其肺组织病变不同程度地改善;其肺泡灌洗液中IL-1β含量和白细胞计数，肺组织中NLRP3蛋白（清肺保元胶囊低剂量组除外）和Cleaved caspase-1蛋白（清肺保元胶囊中、低剂量组除外）的相对表达量均显著降低（P<0.05或P<0.01）。结论：清肺保元胶囊可减轻COPD模型大鼠的肺组织病理损伤、改善其肺功能，这种作用可能与抑制NLRP3信号通路进而抑制炎症反应有关。

关键词 清肺保元胶囊;慢性阻塞性肺疾病;气道炎症;NOD样受体蛋白3信号通路;大鼠

中图分类号 R285.5 文献标志码 A 文章编号 1001-0408（2021）03-0309-05

DOI 10.6039/j.issn.1001-0408.2021.03.10

ABSTRACT   OBJECTIVE： To investigate inhibitory effects of Qingfei baoyuan capsule on airway inflammation in rats with chronic obstructive pulmonary disease （COPD）， and its effects on NLRP3 signaling pathway. METHODS： Totally 60 SD male rats were randomly divided into blank control group， model group， dexamethasone group （positive control， 0.2 mg/kg）， Qingfei baoyuan capsule high-dose， medium-dose and low-dose groups （1 232.0， 616.0， 308.0 mg/kg）， with 10 rats in each group. Except for blank control group， other groups were fumigated for 28 days and given intratracheal dripping of lipopolysaccharide twice to induce COPD model. Since the 29th day after modeling， blank control group and model group were given constant volume of normal saline intragastrically， and administration groups were given related medicine intragastrically. The administration volume was 10 mL/kg， once a day， for consecutive 28 days. After last administration， the lung function was detected. The pathological changes of lung tissue were observed by HE staining. The content of interleukin-1β （IL-1β） in bronchoalveolar lavage fluid （BALF） were detected by ELISA， and the number of leukocytes was counted; the expression of NLRP3 and Cleaved caspase-1 in lung tissue of rats were detected by Western blotting assay. RESULTS： Three， two， one， one and two rats died in model group， dexamethasone group， Qingfei baoyuan capsule high-dose， medium-dose and low-dose groups， respectively. Compared with blank control group， FEV0.3/FVC of rats in model group was significantly decreased （P<0.01）. A large number of inflammatory cells infiltration were found in the lung tissue， and lung tissue lesion was obvious. The content of IL-1β and white blood cell count in BALF， relative expression of NLRP3 and Cleaved caspase-1 protein in lung tissue were increased significantly  （P<0.05 or P<0.01）. Compared with model group， FEV0.3/FVC of administration groups were increased significantly （P<0.05 or P<0.01）; lung tissue lesion of them were improved to different extents. The content of IL-1β and white cell count in BALF， relative expression of NLRP3 protein （except for Qingfei baoyuan capsule low-dose group） and Cleaved caspase-1 protein （except for Qingfei baoyuan capsule medium-dose and low-dose groups） in lung tissue were decreased significantly （P<0.05 or P<0.01）. CONCLUSIONS： Qingfei baoyuan capsule can relieve lung tissue lesion and improve lung function in COPD model rats， the effects of which may be associated with inhibiting inflammation reaction by inhibiting NLRP3 signaling pathway.

KEYWORDS   Qingfei baoyuan capsule; Chronic obstructive pulmonary disease; Airway inflammation; NLRP3 signaling pathway; Rat

慢性阻塞性肺疾病（Chronic obstructive pulmonary disease，COPD），因其高发病率和病死率，已成为全球重大的健康问题[1]。吸烟是COPD重要的危险因素，由于吸烟引起的慢性气道炎症进一步加重了小气道异常和实质损伤[2-3]。近年许多研究表明，NOD样受体蛋白3（NLRP3）通路参与了COPD的气道炎症进程[4-5]，故其已成为COPD治疗的新靶点。……